Development of a Robust Bioprocess to Rapidly Produce Griffithsin, a Broad-Spectrum Therapeutic and Preventive Antiviral Candidate

National Center For Advancing Translational Sciences

Investigators

Linked publications, trials & patents

Abstract

Development of a high-yield E. coli clone was successfully completed, using standard, commercially available BL21 strains. These strains were selected to allow full freedom to operate on any future development of the system. The yields of soluble Q-GRFT and GRFT were confirmed in different scales of expression, including 200-ml, 2-liter and 20-liter cultures, and would be suitable for larger-scale commercial production. The selected clone will be used to make the master cell bank and to continue further upstream and downstream process development. The ongoing upstream development is focused on a bioreactor feeding strategy to achieve an appropriately high cell density (e.g., >80 OD600 at induction). For downstream development, a simplified lab-scale purification strategy was successfully developed using two-column chromatography. The preliminary data showed that the current process yielded 98% pure Q-GRFT and endotoxin was below the detectable limit. Demonstration of the optimized process at pilot scale (20-liter) is ongoing.